Linked Life Data

15576946

Source:http://linkedlifedata.com/resource/pubmed/id/15576946

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:dateCreated
2004-12-3
pubmed:abstractText
The identification of protein binding partners often facilitates understanding of protein complex function. However, identifying binding partners has proven difficult because proteins are often bound to insoluble structures or are only present during certain stages of the cell cycle. Fortunately, Xenopus eggs stockpile many proteins, which are typically insoluble, as soluble subcomplexes to facilitate rapid early embryonic divisions. We exploited this by developing a purification scheme using Xenopus egg extracts to isolate Coomassie-stainable amounts of the xNdc80 kinetochore complex. In this scheme Xenopus eggs are directly made into a mitotic high-speed supernatant and then flowed over three chromatographic columns: heparin, Mono-Q, and Superose 6 gel filtration columns. A final immunoprecipitation is then performed from the peak Superose 6 column to yield Ndc80 complex purified to homogeneity. With minor modification and manipulation of Xenopus egg extracts, this protocol can easily be adapted for purification of other protein complexes.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:issn
1064-3745
pubmed:author
pubmed:issnType
Print
pubmed:volume
296
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
383-91
pubmed:dateRevised
2005-11-17
pubmed:meshHeading
pubmed:year
2005
pubmed:articleTitle
Purification of the Ndc80 kinetochore subcomplex from Xenopus eggs.
pubmed:affiliation
Department of Biochemistry and Molecular Genetics, University of Virginia Medical School, Charlottesville, VA, USA.
pubmed:publicationType
Journal Article